Probe set filtering increases correlation between Affymetrix GeneChip and qRT-PCR expression measurements.

Probe set filtering increases correlation between Affymetrix GeneChip and qRT-PCR expression measurements.
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DOI:
10.1186/1471-2105-11-104
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发表时间:
2010-02-24
期刊:
影响因子:
3
通讯作者:
Pokarowski P
Pokarowski P
中科院分区:
生物学4区
文献类型:
--
作者:
Mieczkowski J;Tyburczy ME;Dabrowski M;Pokarowski P

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Affymetrix GeneChip 微阵列是两种类型研究中表达谱分析的流行平台:检测通过 t 检验的 p 值计算的差异表达,以及估计分析组之间的倍数变化。有许多不同的预处理算法可用于汇总 Affymetrix 数据。这些方法的主要目标是消除非特异性杂交的影响,并最佳地组合来自注释到同一转录本的多个探针的信息。通过与定量逆转录 PCR (qRT-PCR) 等参考方法进行比较,对这些方法进行基准测试。我们对 Affymetrix GeneChip 和 qRT-PCR 结果之间的一致性进行了全面分析。我们分析了 J.N. 引入的分数过滤的影响。 McClintick 和 H.J. Edenberg (2006) 和 2 个映射程序:由 Dai 等人提出的更新的探针集定义。 (2005)和我们的“朴素映射”方法。由于自微阵列设计以来基因组序列注释的演变,我们还研究了注释发布日期的影响。这些比较是针对上述 2 种研究类型中的 6 种流行预处理算法(MAS5、PLIER、RMA、GC-RMA、MBEI 和 MBEImm)而准备的。我们使用来自 6 个独立生物实验的数据集。作为微阵列和 qRT-PCR 值再现性的衡量标准,我们使用线性相关系数和秩相关系数。我们表明,按分数过滤 Present 调用增加了所有 6 种预处理算法的相关性。我们观察到 PM-MM 和仅 PM 方法的性能差异:使用 MM 探针增加了倍数变化研究中的相关性,但事实证明,仅 PM 方法在检测差异表达方面表现更好。我们建议使用 GC-RMA 检测差异表达,使用 PLIER 估计倍数变化。使用更新的注释可以改善两种类型研究的结果,鼓励对旧数据的重新分析。
Affymetrix GeneChip microarrays are popular platforms for expression profiling in two types of studies: detection of differential expression computed by p-values of t-test and estimation of fold change between analyzed groups. There are many different preprocessing algorithms for summarizing Affymetrix data. The main goal of these methods is to remove effects of non-specific hybridization, and to optimally combine information from multiple probes annotated to the same transcript. The methods are benchmarked by comparison with reference methods, such as quantitative reverse-transcription PCR (qRT-PCR). We present a comprehensive analysis of agreement between Affymetrix GeneChip and qRT-PCR results. We analyzed the influence of filtering by fraction Present calls introduced by J.N. McClintick and H.J. Edenberg (2006) and 2 mapping procedures: updated probe sets definitions proposed by Dai et al. (2005) and our "naive mapping" method. Because of evolution of genome sequence annotations since the time when microarrays were designed, we also studied the effect of the annotation release date. These comparisons were prepared for 6 popular preprocessing algorithms (MAS5, PLIER, RMA, GC-RMA, MBEI, and MBEImm) in the 2 above-mentioned types of studies. We used data sets from 6 independent biological experiments. As a measure of reproducibility of microarray and qRT-PCR values, we used linear and rank correlation coefficients. We show that filtering by fraction Present calls increased correlations for all 6 preprocessing algorithms. We observed the difference in performance of PM-MM and PM-only methods: using MM probes increased correlations in fold change studies, but PM-only methods proved to perform better in detection of differential expression. We recommend using GC-RMA for detection of differential expression and PLIER for estimation of fold change. The use of the more recent annotation improves the results in both types of studies, encouraging re-analysis of old data.
DOI: 10.1073/pnas.011404098
发表时间: 2001-01-02
影响因子: 11.1
作者:
Li, C;Wong, WH
通讯作者: Wong, WH
使用更新的探针集定义提高了微阵列的精度和精度。
DOI: 10.1186/1471-2105-8-48
发表时间: 2007-02-08
期刊: BMC bioinformatics
影响因子: 3
作者:
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如何决定?从短寡核苷酸阵列数据中计算基因表达的不同方法将产生不同的结果。
DOI: 10.1186/1471-2105-7-137
发表时间: 2006-03-15
期刊: BMC BIOINFORMATICS
影响因子: 3
作者:
Millenaar, FF;Okyere, J;May, ST;van Zanten, M;Voesenek, LACJ;Peeters, AJM
通讯作者: Peeters, AJM
通过当前调用过滤对微阵列实验分析的影响。
DOI: 10.1186/1471-2105-7-49
发表时间: 2006-01-31
期刊: BMC BIOINFORMATICS
影响因子: 3
作者:
McClintick, JN;Edenberg, HJ
通讯作者: Edenberg, HJ
DOI: 10.1186/gb-2004-5-10-r80
发表时间: 2004
期刊: Genome biology
影响因子: 12.3
作者:
Gentleman RC;Carey VJ;Bates DM;Bolstad B;Dettling M;Dudoit S;Ellis B;Gautier L;Ge Y;Gentry J;Hornik K;Hothorn T;Huber W;Iacus S;Irizarry R;Leisch F;Li C;Maechler M;Rossini AJ;Sawitzki G;Smith C;Smyth G;Tierney L;Yang JY;Zhang J
通讯作者: Zhang J